		Technical Data
G8170-47	Glycogen Synthase Kinase 3, phosphorylated (Y279/Y216) (GSK3)

Description:
Glycogen Synthase Kinase-3 (GSK-3) is a serine/threonine kinase that was first characterized by its role in intermediary metabolism. The enzyme is the mammalian homolog of the zeste-white3 (shaggy) homeotic gene of Drosophila melanogaster and has been implicated in the regulation of the c-Jun/AP-1 transcription factor. Genetic and biochemical evidence now indicates that GSK-3 plays a role in intracellular signaling pathways. For example, during insulin and growth factor signaling the GSK-3 beta isoform becomes phosphorylated on its N-terminal serine-9 residue, presumably by either the action of MAPKAP Kinase-1 (p90 S6 kinase, Rsk-2) or p70 S6 kinase. Such phosphorylation of GSK-3 blocks its ability to phosphorylate c-Jun, protein phosphatase inhibitor 2 or the GSK-3 substrate peptide. GSK-3 is constitutively phosphorylated at this tyrosine residue in resting cells. This immunogen is identical in 15 of 17 residues in human and rat GSK-3a (Y279) and GSK-3b (Y216). The phosphotyrosine residue can be removed from GSK-3 by treatment with PTP1B.

Applications:
Suitable for use in Western Blot. Not suitable for use in Immunoprecipitation. Other applications have not been tested.

Recommended Dilutions:
Western Blot: 0.5-2ug/ml detects phospho-GSK-3 in RIPA lysates from human Jurkat cells. Previous lots detected phospho-GSK-3 in mouse 3T3/A31, human A431 and rat L6 cell lysates. Jurkat cell lysate was resolved by electrophoresis, transferred to nitrocellulose and probed with G8170-47. Proteins were visualized using an IgG (HRP) goat anti-mouse and a chemiluminescence detection system
Optimal dilutions to be determined by the researcher.

Included Positive Antigen Control:
J9000: Jurkat lysate. Use 20ug per lane for minigels.

Storage and Stability:
May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Aliquots are stable for 12 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.

Type	Isotype	Clone	Grade
Mab	IgG1,k	0.T.57	Affinity Purified

Size		Storage	Shipping	Source	Host
200ug		-20°C	Blue Ice	Drosophila	Mouse

Concentration:

~1mg/ml

Immunogen:

Synthetic peptide (aa203-219, C-KQLLHGEPNVS[ pY]ICSRY) corresponding to the catalytic domain (between subdomain VII & VIII) of Drosophila GSK-3/shaggy enzyme coupled to ovalbumin.

Purity:

Purified by Protein G affinity chromatography.

Form

Supplied as a liquid in PBS, pH 7.6, 0.1% sodium azide.

Specificity:

Recognizes Drosophila Glycogen Synthase Kinase 3, phosphorylated (Y279/Y216) (phosphoGSK-3) at 46kD (alpha) and 51kD (beta). Species crossreactivity: mouse, rat, human, S. pombe and Dictyostelium. Expected to crossreact with nearly all species because of the extremely high conservation of this region among the known enzymes of many different species.

Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.

1. Hughes K, et al. 1992. Eur. J. Biochem. 203:305-311. 2. Siddle K, et al. 1995. Biochem. J. 305:25-28. 3. Woodgett JR, et al. 1993. EMBO. 12:803-808. 4. Harwood AJ, et al. 1995. Cell. 80:139-148.